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HPLC法直接同时测定大鼠毛发中的胱氨酸、酪氨酸及组氨酸
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李莉,万子彦,周颖,张闯,崔红艳,肖男,钟家棋,宗豪,吴真,袁凤鸣,秦雅茹,张明 南京医科大学康达学院 
基金项目:江苏省高等学校自然科学研究面上项目(17KJD150005);连云港市科技计划项目(SH1627);南京医科大学科技发展基金项目(2016NJMU011,2016NJMUZD021);南京医科大学康达学院科研发展基金项目(KD2016GCCRCYJ01,KD2016GCCRCYJ02,KD2016KYJJZD001,KD2016KYJJZD002);江苏省高等学校大学生创新创业训练计划(201713980006X)
中文摘要:建立了同时测定大鼠毛发中胱氨酸、酪氨酸和组氨酸的高效液相色谱(HPLC)检测方法。优化了色谱分离及检测条件,样品经酸解后进样分析。采用Titank C18色谱柱(4.6 mm×250 mm,5 μm),以10 mmol/L磷酸氢二铵缓冲液(含10 mmol/L 1-辛烷磺酸钠,用磷酸调至pH 2.0)-乙腈为流动相,进行梯度洗脱,流速1.0 mL/min,检测波长205 nm,进样体积为100 μL,柱温为8 ℃。结果表明,在优化条件下,大鼠毛发中的胱氨酸、酪氨酸和组氨酸能实现有效分离。3种氨基酸的线性良好,相关系数(r)均大于0.999;检出限(S/N=3)为49~442 μg/L;定量下限(S/N=10)为148~884 μg/L;样品的平均加标回收率为97.8%~102%,相对标准偏差(RSD)为0.1%~1.6%。该方法操作简便、准确可靠、重现性好,可用于大鼠毛发中胱氨酸、酪氨酸和组氨酸的检测。
中文关键词:大鼠毛发  胱氨酸  酪氨酸  组氨酸  高效液相色谱(HPLC)
 
Direct and Simultaneous Determination of Cystine,Tyrosine and Histidine in Hair of Rat by HPLC
Abstract:A high performance liquid chromatographic(HPLC) method was developed for the direct and simultaneous determination of cystine,tyrosine and histidine in hair of rat.The chromatographic separation and detection conditions were optimized,and the samples were analyzed after acid hydrolysis.The determination was performed on a Titank C18 column(4.6 mm×250 mm,5 μm) with acetonitrile-10 mmol/L diammonium hydrogen phosphate buffer(containing 10 mmol/L sodium 1-octanesulfonate and obtaining pH 2.0 by phosphoric acid) as mobile phases by gradient elution at a flow rate of 1.0 mL/min.The detection wavelength was set at 205 nm,and the injection volume was 100 μL.The column temperature was set at 8 ℃.Under the optimal conditions,cystine,tyrosine and histidine in hair of rat were effectively separated.Good linearities for the analytes were obtained in their respective ranges,with correlation coefficients(r) larger than 0.999.The limits of detection(S/N=3) were in the range of 49- 442 μg/L and the limits of quantitation(S/N=10) were 148-884 μg/L.The spiked recoveries were in the range of 97.8%-102% with relative standard deviations of 0.1%-1.6%.This method is simple,accurate,reliable and reproducible,and could be applied in the determination of cystine,tyrosine and histidine in hair of rat.
Key Words:hair of rat  cystine  tyrosine  histidine  high performance liquid chromatography(HPLC)
引用本文:李莉,万子彦,周颖,张闯,崔红艳,肖男,钟家棋,宗豪,吴真,袁凤鸣,秦雅茹,张明.HPLC法直接同时测定大鼠毛发中的胱氨酸、酪氨酸及组氨酸[J].分析测试学报,2018,37(12):1485-1489.
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